title: Coculturing cumulus oocyte complexes with denuded oocytes alters zona pellucida ultrastructure in in vitro matured bovine oocytes.
abstract: Oocyte quality is a key factor affecting success of in vitro embryo production in cattle. Improving the microenvironment of oocytes during in vitro maturation (IVM) can increase developmental rate and embryo quality. Therefore, the objective was to determine whether denuded oocytes (DO) affect embryo development and ultrastructure of the zona pellucida (ZP) in in vitro matured bovine oocytes. Intact immature cumulus-oocytes complexes (COC) obtained from a local abattoir or by ovum pick-up (OPU) were cocultured with and without abattoir-obtained DO at a COC:DO ratio of 1:5. After IVM, DO were removed and intact DO were either fertilized or observed by scanning electron microscopy. Blastocyst quality was evaluated using a TUNEL assay. The ZP pore size decreased after IVM in COC + DO coculture, regardless of their origin (OPU, 310.5 ± 92.5 vs. 428.9 ± 148.5 nm; abattoir, 317.5 ± 68.5 vs. 358.9 ± 128.5 nm; P < 0.05; mean values ± standard deviation). Moreover, the number of ZP pores in OPU COC + DO and COC + DO was greater than those in OPU COC and COC (control) groups (56 ± 4 and 55 ± 7 vs. 50 ± 6 and 42 ± 4; P < 0.05). The rate of blastocyst development in COC + DO and OPU COC + DO groups was greater those in control and OPU COC groups (36.6% and 55.5% vs. 28.1% and 40.0%; P < 0.05). Moreover, the total cell numbers of blastocysts in COC + DO group exceeded that of control (132.91 ± 30.90 vs. 115.44 ± 24.95; P < 0.05), with no significant between OPU COC + DO and OPU COC groups (139.31 ± 42.51 vs. 137.00 ± 61.34). In conclusion, in vitro embryo development competence and quality improved when oocytes were cocultured with DO. Furthermore, there more, but smaller, ZP pores.
authors.affiliation: Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University, Jinju, Republic of Korea.
authors.forename: Byung-Hyun
authors.initials: BH
authors.lastname: Choi
authors.forename: Jae-Il
authors.initials: JI
authors.lastname: Bang
authors.forename: Jong-In
authors.initials: JI
authors.lastname: Jin
authors.forename: Seong-Su
authors.initials: SS
authors.lastname: Kim
authors.forename: Hyun-Tae
authors.initials: HT
authors.lastname: Jo
authors.forename: Gautam Kumar
authors.initials: GK
authors.lastname: Deb
authors.forename: Nasser
authors.initials: N
authors.lastname: Ghanem
authors.forename: Kyu-Woan
authors.initials: KW
authors.lastname: Cho
authors.forename: Il-Keun
authors.initials: IK
authors.lastname: Kong
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology / Q000648:ultrastructure; D002417:Cattle; D018920:Coculture Techniques / Q000662:veterinary*; D054885:Cumulus Cells / Q000648:ultrastructure; D046149:Embryo Culture Techniques / Q000662:veterinary*; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods / Q000662:veterinary*; D008855:Microscopy, Electron, Scanning; D054315:Oocyte Retrieval / Q000662:veterinary; D009865:Oocytes / Q000166:cytology / Q000648:ultrastructure; D015044:Zona Pellucida / Q000648:ultrastructure*
keywords: Apoptosis; Bovine oocytes; Coculture; Embryo development; Zona pellucida